Integrated liquid chromatography-multiangle light scattering system to measure absolute masses and stoichiometries of biological assemblies in solution

集成液相色谱-多角度光散射系统，用于测量溶液中生物组件的绝对质量和化学计量

• 批准号: RTI-2019-00728
• 负责人: Melacini, Giuseppe
• 金额: $ 10.92万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=659504
• 关键词: Integrated; liquid; chromatography; multiangle; light

This application requests a state-of-the-art size exclusion chromatography system integrated with multi-angle light scattering (SEC-MALS). SEC-MALS is capable of measuring absolute aggregate molecular weights (MWs) and radii of gyration (Rg) under physiological native solution conditions with unparalleled accuracy, precision and throughput not attainable by conventional analytical SEC. Reliable MW and Rg values are needed to determine the oligomerization state and binding stoichiometry of complexes formed by a wide range of proteins and polymers at the center of a multitude of research programs at McMaster. These include not only globular proteins, but also membrane proteins, intrinsically disordered proteins, non-fouling polymers as well as amyloidogenic polypeptides, which are notoriously refractory to other methods for measuring MW and Rg. The requested instrument, a Wyatt MALS-RI with GE Healthcare SEC, is one of the most versatile and reliable SEC-MALS configurations for academic work currently on the market and will have a transformative impact on a collaborative community of > 8 different NSERC-funded research groups spread across three different faculties at McMaster university, i.e. Faculty of Science, Faculty of Health Sciences and Faculty of Engineering. Currently, there is no SEC-MALS at McMaster. Given the outstanding performance in terms of accuracy, sensitivity and throughput, SEC-MALS is now the gold standard requested by the best international journals for the accurate determination of oligomeric states and macromolecular complex stoichiometries. Without SEC-MALS it is becoming nearly impossible to publish high-impact papers. Furthermore, SEC-MALS has been adopted as the industry standard for the characterization of higher-order structures formed by therapeutic biologicals and polymers. Hence, SEC-MALS will dramatically enhance our ability to train the next generation of HQP in preparation for jobs not only in the academia, but also in the pharma and biotech-sectors. The absence of SEC-MALS at McMaster is severely limiting dozens of researchers and is jeopardizing their ability to maintain internationally competitive research programs. If funded, the requested instrument will be the first SEC-MALS in the whole McMaster campus and will be installed in a newly-renovated centralized, shared and widely accessible core facility. We anticipate an extremely high demand by all groups working in the structural biology, enzymology and bioengineering fields. For virtually all projects in these fields, it is imperative to reliably establish the stoichiometry of homo- and hetero-oligomers before further progress can be achieved and currently SEC-MALS is the prime technique for this purpose. The impact of the research enabled by the requested SEC-MALS ranges from the understanding of the mechanisms of allosteric regulation to the design of new biomaterials. Given the absence of SEC-MALS at McMaster, SEC-MALS is a clear and urgent need. **

该应用需要集成多角度光散射 (SEC-MALS) 的最先进的尺寸排阻色谱系统。 SEC-MALS 能够在生理天然溶液条件下测量绝对聚集分子量 (MW) 和回转半径 (Rg)，具有传统分析 SEC 无法达到的无与伦比的准确性、精密度和通量。需要可靠的 MW 和 Rg 值来确定麦克马斯特众多研究项目中心的各种蛋白质和聚合物形成的复合物的寡聚状态和结合化学计量。这些不仅包括球状蛋白，还包括膜蛋白、本质无序蛋白、防污聚合物以及淀粉样蛋白多肽，众所周知，这些蛋白对其他测量 MW 和 Rg 的方法是无效的。所请求的仪器是带有 GE Healthcare SEC 的 Wyatt MALS-RI，是目前市场上用于学术工作的最通用、最可靠的 SEC-MALS 配置之一，将对超过 8 个不同 NSERC 资助的协作社区产生变革性影响研究小组分布在麦克马斯特大学的三个不同院系，即理学院、健康科学学院和工程学院。 目前，麦克马斯特大学没有 SEC-MALS。 鉴于在准确性、灵敏度和通量方面的出色表现，SEC-MALS 现已成为最佳国际期刊要求的准确测定低聚状态和大分子复杂化学计量的黄金标准。如果没有 SEC-MALS，几乎不可能发表高影响力的论文。此外，SEC-MALS 已被采用作为表征治疗性生物制品和聚合物形成的高阶结构的行业标准。因此，SEC-MALS 将极大地提高我们培训下一代 HQP 的能力，为学术界、制药和生物技术领域的工作做好准备。麦克马斯特大学缺乏 SEC-MALS 严重限制了数十名研究人员的工作，并危及他们维持具有国际竞争力的研究项目的能力。 如果获得资助，所需的仪器将成为整个麦克马斯特校区的第一个 SEC-MALS，并将安装在新装修的集中、共享且可广泛访问的核心设施中。我们预计结构生物学、酶学和生物工程领域的所有团队都会提出极高的需求。 对于这些领域的几乎所有项目，在取得进一步进展之前，必须可靠地建立同源和异源低聚物的化学计量，目前 SEC-MALS 是实现此目的的主要技术。所要求的 SEC-MALS 所实现的研究影响范围从对变构调节机制的理解到新生物材料的设计。鉴于麦克马斯特大学缺乏 SEC-MALS，显然迫切需要 SEC-MALS。 **